本文選題：鼻咽癌 + 骨髓間充質(zhì)干細胞�。� 參考：《中華腫瘤防治雜志》2014年23期

【摘要】：目的初步探討人骨髓間充質(zhì)干細胞(human bone marrow mesenchymal stem cells,hbMSCs)條件培養(yǎng)基(conditioned medium,CM)在人鼻咽癌CNE1及CNE2細胞上皮間質(zhì)轉(zhuǎn)化(epithelial mesenchymal transition,EMT)中的影響。方法取第3代hbMSCs融合達70%~80%時,換液培養(yǎng)24h后取其上清液(hbMSCs-CM);取對數(shù)生長期5×103CNE1及CNE2細胞,分別在普通培養(yǎng)液(對照組)和hbMSCs-CM的培養(yǎng)液(實驗組)中培養(yǎng)。使用倒置相差顯微鏡下觀察細胞形態(tài)學改變,蛋白質(zhì)印跡法及q-PCR檢測EMT相關(guān)蛋白及RNA變化。結(jié)果 hbMSCs-CM培養(yǎng)鼻咽癌細胞5d,CNE1和CNE2均觀察到細胞間距離增寬,并且折光性強、多量單極或雙極梭形狀成纖維細胞樣細胞,以CNE2細胞最為明顯。q-PCR檢測結(jié)果顯示,與對照組相比,hbMSCs-CM培養(yǎng)組細胞上皮細胞標志基因mRNA的相對表達水平下調(diào),差異有統(tǒng)計學意義,P值均0.001;間質(zhì)細胞標志基因mRNA相對表達水平上調(diào),差異有統(tǒng)計學意義,P值均0.001;EMT相關(guān)轉(zhuǎn)錄因子基因mRNA相對表達量上調(diào),差異有統(tǒng)計學意義,P值均0.001。蛋白質(zhì)印跡法檢測結(jié)果顯示,hbMSCs-CM誘導CNE1及CNE2細胞發(fā)生EMT并呈一定濃度依賴性。當hbMSCs-CM濃度≥70%,上皮細胞標志蛋白相對表達水平顯著下降,差異有統(tǒng)計學意義,P0.05。當hbMSCs-CM濃度50%時,CNE1和CNE2細胞間質(zhì)細胞標志及轉(zhuǎn)錄因子蛋白相對表達水平上調(diào),差異有統(tǒng)計學意義,P0.05。結(jié)論 hbMSCs可改變鼻咽癌細胞形態(tài),并下調(diào)細胞黏附蛋白及增加細胞間質(zhì)蛋白水平,可能是參與鼻咽癌上皮間質(zhì)轉(zhuǎn)化過程的因素之一。
[Abstract]:Objective to investigate the effect of human bone marrow mesenchymal stem cells (hbMSCs) conditioned mediums CM) on epithelial mesenchymal transition of human nasopharyngeal carcinoma (CNE1) and CNE2 cells (CNE2 cells). Methods\\\; Methods when the third generation of hbMSCs fused to 70 ~ 80, the supernatant of HbMSCs-CMN was cultured for 24 hours, and the 5 脳 103CNE1 and CNE2 cells in logarithmic growth period were cultured in normal medium (control group) and hbMSCs-CM medium (experimental group), respectively. Morphologic changes were observed under inverted phase contrast microscope, EMT related proteins and RNA were detected by Western blot and q-PCR. Results for 5 days, hbMSCs-CM cultured nasopharyngeal carcinoma cell line CNE1 and CNE2 showed that the distance between cells was widened and the refractive index was strong. The results of multipolar or bipolar fusiform fibroblast-like cells were most obvious in CNE2 cells. Compared with the control group, the relative expression level of epithelial marker gene mRNA in hbMSCs-CM culture group was down-regulated, the difference was significant (P = 0.001), and the relative expression level of interstitial cell marker gene mRNA was up-regulated. The difference was statistically significant (P = 0.001). The relative expression of transcription factor gene mRNA was up-regulated, and the difference was statistically significant (P = 0.001). Western blot analysis showed that hbMSCs-CM induced EMT in CNE1 and CNE2 cells in a dose-dependent manner. When the concentration of hbMSCs-CM was 鈮，

本文編號：1801451